TRANSEPITHELIAL TRANSPORT AND ENZYMATIC DETOXIFICATION OF GLUTEN IN MACAQUES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们的特点是条件面筋敏感性在青少年恒河猴，是在许多方面类似于人类条件面筋敏感性，腹腔疾病。 因此，这种谷蛋白敏感性的动物模型可能有助于研究乳糜泻的发病机制和治疗。在这里，我们进行了两个试点实验，以证明该模型的潜在效用，用于研究肠道通透性对免疫毒性谷蛋白肽和药理解毒的面筋在体内的口服酶药物候选人。在年龄匹配的谷蛋白敏感性和对照猕猴中，通过使用质谱法检测和定量口服给药的同位素标记的33聚体谷蛋白肽穿过肠上皮传递到血浆，研究了肠通透性。 在谷蛋白敏感的猕猴中，通过在有或没有EP-B2补充的情况下每天给予谷蛋白挑战，评价了治疗上有前景的口服蛋白酶EP-B2的保护作用。 对这只猕猴和年龄匹配的对照进行了基于ELISA的抗体测定和盲法临床评价，以评估对谷蛋白的反应。标记的33-mer肽检测在血浆中的谷蛋白敏感的猕猴，无论是在缓解期和活动期的疾病，但不是在健康对照组的血浆。 施用EP-B2而不是媒介物预防了响应于饮食麸质挑战的临床复发。出乎意料的是，在EP-B2治疗阶段观察到抗麦醇溶蛋白（IgG和伊加）和抗组织转氨酶（IgG）抗体显著增加。 面筋敏感的恒河猴可能是一个有吸引力的资源，研究腹腔疾病的重要方面，包括增强肠道通透性和口服酶药物候选人的药理学。口服剂量的EP-B2对摄入的麸质发挥保护作用。 有限的数据表明，由胃活性谷蛋白酶产生的短谷蛋白肽的渗透性增强可能引发抗体应答升高，但这些抗体不一定是临床疾病的病因。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We characterized a condition of gluten sensitivity in juvenile rhesus macaques that is similar in many respects to the human condition of gluten sensitivity, celiac disease. This animal model of gluten sensitivity may therefore be useful toward studying both the pathogenesis and the treatment of celiac disease. Here, we perform two pilot experiments to demonstrate the potential utility of this model for studying intestinal permeability toward an immunotoxic gluten peptide and pharmacological detoxification of gluten in vivo by an oral enzyme drug candidate. Intestinal permeability was investigated in age-matched gluten-sensitive and control macaques by using mass spectrometry to detect and quantify an orally dosed, isotope labeled 33-mer gluten peptide delivered across the intestinal epithelium to the plasma. The protective effect of a therapeutically promising oral protease, EP-B2, was evaluated in a gluten-sensitive macaque by administering a daily gluten challenge with or without EP-B2 supplementation. ELISA-based antibody assays and blinded clinical evaluations of this macaque and of an age-matched control were conducted to assess responses to gluten. Labeled 33-mer peptide was detected in the plasma of a gluten-sensitive macaque, both in remission and during active disease, but not in the plasma of healthy controls. Administration of EP-B2, but not vehicle, prevented clinical relapse in response to a dietary gluten challenge. Unexpectedly, a marked increase in anti-gliadin (IgG and IgA) and anti-tissue transglutaminase (IgG) antibodies was observed during the EP-B2 treatment phase. Gluten-sensitive rhesus macaques may be an attractive resource for investigating important aspects of celiac disease, including enhanced intestinal permeability and pharmacology of oral enzyme drug candidates. Orally dosed EP-B2 exerts a protective effect against ingested gluten. Limited data suggest that enhanced permeability of short gluten peptides generated by gastrically active glutenases may trigger an elevated antibody response, but that these antibodies are not necessarily causative of clinical illness.